Phage-display technology has begun to make critical contributions to the study of molecular recognition. DNA sequences are cloned into phage, which then present on their surface the proteins encoded by the DNA. Individual phage are rescued through interaction of the displayed protein with a ligand, and the specific phage is amplified by infection of bacteria.
Phage-display technology is powerful but challenging and the aim of this manual is to provide comprehensive instruction in its theoretical and applied so that any scientist with even modest molecular biology experience can effectively employ it. The manual reflects nearly a decade of experience with students of greatly varying technical expertise andexperience who attended a course on the technology at Cold Spring Harbor Laboratory.
Phage-display technology is growing in importance and power. This manual is an unrivalled source of expertise in its execution and application.
"The book's raison d'être is as a source of detailed protocols. These are clearly written, and include ideas for experimental controls and useful trouble-shooting notes. Jargon is generally avoided, and the protocols are up to date.
[This] is a much-needed and well-executed attempt at theoretical and practical support to any researcher, established or novice, using phage display."
Trends in Genetics
"This book is a solid work that addresses both theoretical and practical aspects of phage display technology. It covers the use of phage display in protein engineering with particular emphasis on antibody, peptide, cDNA libraries and ligand receptor engineering, which together represent [approximately] 90% of the phage display field. Anyone using or contemplating any of these applications would find it worthwhile to read the relevant chapters."
Trends in Biotechnology
- SECTION 1 PHAGE DISPLAY
- Chapter 1. Filamentous Phage Biology
- Chapter 2. Phage-display Vectors
- Chapter 3. Antibody Libraries
- Chapter 4. Peptide Libraries
- Chapter 5. Functional Domains and Scaffolds
- Chapter 6. Gene Fragment Libraries and Genomic and cDNA Expression Cloning
- SECTION 2 ANTIBODY LIBRARIES
- Chapter 7. Overview: Amplification of Antibody Genes
- Chapter 8. Generation of Antibody Libraries: Immunization, RNA Preparation, and cDNA Synthesis
- Chapter 9. Generation of Antibody Libraries: PCR Amplification and Assembly of Light- And Heavy-chain Coding Sequences
- Chapter 10. Selection from Antibody Libraries
- Chapter 11. Analysis of Selected Antibodies
- Chapter 12. Production and Purification of Fab and scFv
- Chapter 13. Antibody Engineering
- SECTION 3 PEPTIDE LIBRARIES
- Chapter 14. Overview: Peptide Libraries
- Chapter 15. General Phage Methods
- Chapter 16. Production of Peptide Libraries
- Chapter 17. Screening Peptide Libraries
- Chapter 18. Analysis of Phage-borne Peptides
- Chapter 19. Construction and Use of pIII-displayed Peptide Libraries
- SECTION 4 OTHER LIBRARIES AND OTHER METHODS OF PANNING
- Chapter 20. Construction and Selection from Gene Fragment Phage-display Expression Libraries
- Chapter 21. Construction and Selection from cDNA Phage-display Expression Libraries
- Chapter 22. In Vivo Selection of Phage-display Libraries
- Chapter 23. Cell-surface Selection and Analysis of Monoclonal Antibodies from Phage Libraries
- SECTION 5 APPENDICES
- Appendix 1. Useful Information
- Appendix 2. Recipes
- Appendix 3. General Procedures
- Appendix 4. Cautions
- Appendix 5. Suppliers
- Appendix 6. Trademarks